• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.5
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• pp.1386-1390
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• 2005

Lycii Fructus (LF) has been reported to possess various biological activities. In this study, we investigated the anti-inflammatory and anti-allergic effects of LF extract in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells and rat peritoneal mast cells(RPMC). LF extract inhibited NO production and the expressions of inducible NO synthase (iNOS) mRNA and enzyme protein, determined by Griess reactions, RT-PCR and Western blotting, respectively, in LPS-stimulated RAW 264.7 macrophages cells. LF extract significantly inhibited histamine release in compound48/80 stimulated rat peritoneal mast cells compared with the compound48/80 only treated cells. It is suggested that the LF extract can improve the inflammation status that involves the histamine release.

• Herbal Formula Science
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• v.18 no.2
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• pp.183-199
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• 2010

Rheumatoid arthritis is characterized by the focal loss of cartilage due to an up-regulation of inflammatory pathways, which produce inflammatory mediators, such as interleukin-1beta(IL-$1{\beta}$), IL-6, tumour necrosis factor alpha(TNF-$\alpha$), prostaglandin, and nitric oxide(NO). We investigated the anti-arthritic effects of water extract from FLOS LONICERAE(FLWE) in vitro and in vivo. Extract inhibited the production of inflammatory mediators(NO, IL-$1{\beta}$, TNF-$\alpha$, and prostaglandin $E_2$) and the expression of inducible NO synthase(iNOS) and cyclooxygenase-2(COX-2) in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages in a dose-dependent manner. FLWE also inhibited TNF-$\alpha$, IL-$1{\beta}$, IL-6, and $PGE_2$ production as well as COX activity in collagen-induced mouse arthritis. Moreover, FLWE significantly suppressed collagen-induced mouse arthritis. These results suggest that FLOS LONICERAE may be useful for therapy against inflammatory immune diseases and rheumatoid arthritis, probably by suppressing the production of inflammatory mediators.

• The Korean Journal of Food And Nutrition
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• v.34 no.5
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• pp.441-448
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• 2021

This study was designed to investigate the intracellular signaling pathways and immunoenhancing effect of macrophage activation by crude polysaccharides (CPP) extracted from citrus peels. CPP did not affect the cytotoxicity of RAW264.7 cells, but showed dose-dependent effects on cell viability. Also, CPP showed high production of chemokine (nitric oxide (NO)) and cytokines (interleukin (IL)-6 and tumor necrosis factor (TNF)-α). CPP increased IL-6, TNF-α, and inducible nitric oxide synthase (iNOS) mRNA expression dose-dependently. CPP also strongly induced the phosphorylation of the ERK, p38, and IκBα pathways in RAW 264.7 cells. In anti-pattern recognition receptors (PRRs) experiments, the effect of CPP on NO production was strongly suppressed by neutralizing toll-like receptor (TLR)2, TLR4, and Dectin1 antibodies, whereas IL-6 and TNF-α production by CPP was mainly suppressed by mannose receptor (MR). Therefore, these results suggest that CPP treatment-induced NO production was regulated by the ERK, p38, and NF-κB pathways through TLR2, TLR4, and Dectin1 receptors, whereas IL-6 and TNF-α production was primarily regulated by the ERK, p38, and NF-κB pathways through MR receptors.

• Journal of Life Science
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• v.24 no.4
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• pp.370-376
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• 2014

Although the grasshopper Oxya chinensis sinuosa has long been used as food in Korea, there is little data on its functional effects. In this study, we investigated the anti-inflammatory effect of O. c. sinuosa ethanol extract (OCE) in RAW 264.7 mouse macrophage cells treated with lipopolysaccharide (LPS) for induction of inflammation. First, we determined that there is no cytotoxicity at $2,000{\mu}g/ml$ or less of OCE in RAW 264.7 cells. To evaluate the anti-inflammatory effects of OCE, we investigated expression levels of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, and pro-inflammatory enzymes such as inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. In addition, we examined whether OCE could inhibit translocation of NF-${\kappa}B$ p65 into the nucleus in LPS induced RAW 264.7 cells. As a result, we found that the mRNA and protein levels of TNF-${\alpha}$ and IL-6 decreased in LPS-induced RAW 264.7 cells after treatment with OCE in a dose-dependent manner. In addition, we confirmed a $2,000{\mu}g/ml$ concentration of OCE inhibited translocation of NF-${\kappa}B$ p65 by immunnostaining and Western blot analysis, and a decrease in the protein expression levels of iNOS and COX-2. Accordingly, we suppose that OCE has an anti-inflammatory effect through down-regulation of TNF-${\alpha}$, IL-6, iNOS, and COX-2 related to ${\kappa}B$ p65 inflammatory signaling pathways.

• Journal of Life Science
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• v.27 no.6
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• pp.640-645
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• 2017

Clerodendrum trichotomum (CT) leaves and stems have been used in folk medicine for their anti-hypertension, arthritis, rheumatism, and anti-inflammatory properties. This study was performed to evaluate the potential of CT as an anti-oxidant and anti-inflammatory agent. CT leaves were extracted using 70% ethanol (EtOH). Then, using this extract, a hexane, chloroform ($CHCl_3$), ethyl acetate (EtOAc), and n-butanol (BuOH) fraction was prepared. The polyphenol contents were higher in the EtOAc fraction ($78.08{\mu}g/mg$) and BuOH fraction ($77.54{\mu}g/mg$) compared to the other fractions. Also, these two fractions exhibited strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities. Xanthine oxidase inhibitory activities were higher in the $CHCl_3$ fraction ($IC_{50}=4.43{\mu}g/ml$) and EtOAc fraction ($IC_{50}=5.69{\mu}g/ml$). Moreover, the EtOAc fraction effectively inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells ($IC_{50}=18.87{\mu}g/ml$). Thus, we investigated the effects of the EtOAc fraction on the expression of pro-inflammatory cytokines, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) in LPS-stimulated RAW 264.7 cells. The treatment of the EtOAc fraction ($100{\mu}g/ml$) effectively decreased the levels of the tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$) and interleukin-6 (IL-6), and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). These results suggest the potential for CT extract and fractions as promising anti-oxidant and anti-inflammatory agents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.10
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• pp.1527-1534
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• 2014

In this study, we investigated the anti-inflammatory effects of Lactobacillus rhamnosus fermented black garlic (FBG) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. FBG did not show cytotoxicity in RAW 264.7 cells at concentrations less than $800{\mu}g/mL$, and cell viability increased with FBG concentration. Nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production as well as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and IL-6 formation decreased in an FBG concentration-dependent manner, in LPS-induced RAW 264.7 cells. Furthermore, activation of LPS-inducible nitric synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B (NF-${\kappa}B$), and inhibitory kappa B ($I{\kappa}B$) protein expression was effectively inhibited by FBG treatment in LPS-induced RAW 264.7 cells. In contrast, heme oxygenase-1 (HO-1) protein expression significantly increased. These results indicate that the anti-inflammatory activity of FBG was due to activation of NF-${\kappa}B$, inhibition of cytokine production, and expression of iNOS and COX-2. From these results, we expect that FBG could contribute to the prevention and improvement of inflammatory disease.

• Applied Chemistry for Engineering
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• v.30 no.2
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• pp.145-150
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• 2019

In this study, we used extracts obtained from five different solvents (water, ethanol, hexane, ethyl acetate, butanol) of Achyranthes japonica (AJ) and also AJ fermented with Lactobacillus plantarum (LP) to confirm effects on the anti-inflammatory activity in RAW264.7 cells. Experiments of measuring nitric oxide (NO) and cytokine production were performed in lipopolysaccharide (LPS)-induced RAW264.7 cells, and the expression of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was observed by a western blot method. The cytotoxicity of RAW264.7 was confirmed by the cell counting kit (CCK) assay at a concentration of $100{\mu}g/mL$, which has no toxicity. As a result of the inhibition of NO production, the inhibition rate of AJ-LP extracted with ethanol samples was about 74% higher than that of using the control group. Interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and Interleukin-$1{\beta}$ (IL-$1{\beta}$), which are inflammatory cytokines, also showed an excellent efficacy with inhibition rates of about 57, 70, and 74%, respectively. Comparing to the results of COX-2 and iNOS expression in the AJ group, the inhibition rate of 20-hydroxyecdysone was the highest than others. On the other hand, the COX-2 expression level of AJ-LP group decreased about 16% compared to that of the control group, and the iNOS expression level was also decreased about 7%. These results suggest that the extract of AJ fermented from L. plantarum can be used as an anti-inflammatory natural material.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.2
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• pp.310-318
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• 2010

Dioscorea bulbifera is one of the traditional medicinal herb. It commonly used in the treatment of hematemesis, epistaxis, tuberculous cervical lymphadenitis, laryngitis, acute infectious disease in East Asia. In the present study, we have demonstrated the anti-inflammatory effects of Dioscorea bulbifera MeOH extract (DBME) in macrophage cell line. To investigate mechanism of the anti-inflammatory activity, we examined the effects of the lipopolysaccaride (LPS)-induced production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), pro-inflammatory cytokines and expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), p-inhibitory ${\kappa}B{\alpha}$ (p-$I{\kappa}B{\alpha}$), and nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in a murine macrophage cell line RAW 264.7. The RAW 264.7 cells were cultured in DMEM + serum medium for 24 hrs. After serum starvation for 24 hrs, the cells were treated with DBME 0.03, 0.10, 0.30 mg/$m{\ell}$ for 1 h, followed by stimulation with LPS (1 ${\mu}g/m{\ell}$) for activation of immune response. After treatment, cell viability was measured by MTT assay, and NO production was monitored by measuring the nitrite content in culture medium. The protein band of iNOS, COX-2, p-$I{\kappa}B{\alpha}$, and NF-${\kappa}B$ was determined by immunoblot analysis and levels of cytokine were analyzed by sandwich immunoassays. There were three experimental groups: carrageenan, DBME 0.3, 1.0 g/kg. Rats were administrated either carrageenan (40% PEG) or carrageenan + DBME (0.3, 1.0 g/kg body weight) for 4 days (p.o.). To induce acute paw edema, rats were injected 1% carrageenan (100 ${\mu}{\ell}$/rat, dissolved in sterilized saline). The effect of DBME in the carrageenan-induced rat paw edema. As results, DBME has an inhibitory effect on the production of NO, PGE2, TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 and on the expression of iNOS, COX-2, p-$I{\kappa}B{\alpha}$ and translocation of NF-${\kappa}B$ to nuclear from cytosol. In addition, DBME effectively inhibited the increases of paw edema induced by carrageenan treatment in vivo. These results suggest that DBME can inhibit production of pro-inflammatory mediators and might be a useful source for treatment of acute inflammatory disease.

• Korean Journal of Pharmacognosy
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• v.47 no.4
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• pp.327-332
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• 2016

In this study, antioxidant and anti-inflammatory activities of the extract of different organs such as seed, fruit and root from Trichosanthes kirilowii Maxim. (TKM) were investigated in vitro. Among organs of TKM, the methanol extract of seed showed weak 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities with $10.4{\pm}0.1%$ and $25.5{\pm}1.1%$ at $50{\mu}g/mL$ and $100{\mu}g/mL$, respectively, and strong 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) radical scavenging activities with $72.4{\pm}6.3%$ and $96.4{\pm}2.3%$ at $50{\mu}g/mL$ and $100{\mu}g/mL$, respectively. Anti-inflammatory effects of the extracts were investigated by measuring nitric oxide (NO) production, mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced Raw 264.7 macrophage cells. The methanol extracts of seed and fruit at $50{\mu}g/mL$ potently suppressed LPS-stimulated production of NO and reduced the expression of iNOS mRNA. The methanol extracts of seed and fruit also reduced the expression of COX-2 mRNA remarkably. Therefore, seed and fruit of TKM may be utilized for a wide range of health benefits associated with antioxidant and anti-inflammatory activities.

• Nutrition Research and Practice
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• v.14 no.2
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• pp.109-116
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• 2020

BACKGROUND/OBJECTIVES: Excessive intake of simple sugars induces obesity and increases the risk of inflammation. Thus, interest in alternative sweeteners as a sugar substitute is increasing. The purpose of this study was to determine the effect of saccharin on inflammation in 3T3-L1 adipocytes. MATERIALS/METHODS: 3T3-L1 preadipocytes were differentiated into adipocytes. The adipocytes were treated with saccharin (0, 50, 100, and 200 ㎍/mL) for 24 h. Inflammation was induced by exposure of treated adipocytes to lipopolysaccharide (LPS) for 18 h and cell proliferation was measured. The concentration of nitric oxide (NO) was measured by using Griess reagent. Protein expressions of nuclear factor kappa B (NF-κB) and inhibitor κB (IκB) were determined by western blot analysis. The mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin 1β (IL-1β), interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor-α (TNF-α) were determined by real-time PCR. RESULTS: Compared with the control group, the amount of NO and the mRNA expression of iNOS in the LPS-treated group were increased by about 17.6% and 46.9%, respectively, (P < 0.05), and those parameter levels were significantly decreased by saccharin treatment (P < 0.05). Protein expression of NF-κB was decreased and that of IκB was increased by saccharin treatment (P < 0.05). Saccharin decreased the mRNA expression of COX-2 and the inflammation cytokines (IL-1β, IL-6, MCP-1, and TNF-α) (P < 0.05). CONCLUSIONS: The results of this study suggest that saccharin can inhibit LPS-induced inflammatory responses in 3T3-L1 adipocytes via the NF-κB pathway.